Fig. 8.

Postsynaptic BDNF is required for NL1-induced presynaptic maturation. A, B Representative images and quantification of cultured hippocampal neurons from BDNF^lox/lox mice co-transfected with NL1–IRES–mOrange (red) + Cre/GFP (green) on DIV4. Neurons were fixed on DIV6 and immunostained for bassoon (blue) following a 6-h LatA treatment. For separation of the three fluorescent channels, see Additional File 6: Fig. S6 A. Mean ± SEM; N = 3 experiments, n = 10 cells per condition and experiment; two-way ANOVA with post hoc Sidak tests: interaction is significant: ***p< 0.0001; F(1, 116)=259.8; ***p< 0.0001 for NL1 + Cre control vs. NL1 + Cre + LatA; p> 0.05 for NL1 + GFP control vs. NL1 + GFP + LatA. Scale bar is 10 μm. C, D Representative images and quantification of cultured hippocampal neurons from BDNF^lox/lox mice co-transfected with NL1-mOrange (red) + Cre/GFP (green) on DIV4. On DIV6 cultures were stimulated in the presence of antibodies against the lumenal domain of synaptotagmin-1 (Syt1) to label recycling synaptic vesicles. After washing, the cells were fixed and immunolabeled with secondary antibodies against the Syt1 antibody (blue). For separation of the three fluorescent channels, see Additional File 6: Fig. S6 B. Mean ± SEM; N = 3 experiments, n = 10 cells per condition and experiment; Student’s t test: ***p< 0.0001. Scale bar is 10 μm